LOCUS       sequence-366861-        5282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the sleeping chironomid protein 
            pvLEA22mer_shuffle_5. This is one of the top performing targets in 
            our apoptosis assay (it protected from apoptosis)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5282)
  AUTHORS   Veling MT, Nguyen DT, Thadani NN, Oster ME, Rollins NJ, Brock KP, 
            Bethel NP, Lim S, Baker D, Way JC, Marks DS, Chang RL, Silver PA
  TITLE     Natural and Designed Proteins Inspired by Extremotolerant Organisms 
            Can Form Condensates and Attenuate Apoptosis in Human Cells.
  JOURNAL   ACS Synth Biol. 2022 Mar 18;11(3):1292-1302. doi: 
            10.1021/acssynbio.1c00572. Epub 2022 Feb 18.
  PUBMED    35176859
REFERENCE   2  (bases 1 to 5282)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.1c00572"; journalName: "ACS Synth Biol"; date: 
            "2022-03-18- 18"; volume: "11"; issue: "3"; pages: "1292-1302"
COMMENT     Sequence Label: pST_pvLEA22mer_shuffle_5 (pBS0731)
FEATURES             Location/Qualifiers
     source          1..5282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..48)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        220..599
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        600..803
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     754..774
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     848..867
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        848..866
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1275..1292)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1281..1505
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1551..1979
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1638..1657)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1848..1869
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1988..2008)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1993..2322
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2173..2308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2235..2254
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2370..2417
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2436..2810
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    2940..3061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2977..2996)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3031..3050
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3110..3126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3110..3126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3123..3145)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3134..3150
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3158..3164,3165..3182,3183..3188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3341..3358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3512..4100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3592..3611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4271..5062,5063..5131))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4894..4913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5132..5236)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
       61 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
      121 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
      181 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
      241 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      301 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      361 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      421 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      481 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      541 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      601 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      661 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      721 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      781 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
      841 tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagccacca tgacaaaagc
      901 tgagaaggcc ggcgagaagg gagacaaggc cgaggacaag ggcgagacca aggccggatg
      961 atgaattgcc aattgaaatt tgacggccta tgtcactgaa actatccaaa caccccatgt
     1021 cgatactgaa cgtatcgacg catacctcct tccttgaaaa cgcacaatca tacaactggg
     1081 cacataatgc gtacgcccat ctagtacacc catctctgta ggtccagttc aagagctgga
     1141 agagcaccct ccacttagac gtgccgtaag atagaggtta gtgagaatag ccatccaata
     1201 tgagcgttga tagctagacg aggataaggc agtgatagaa ggggcaccaa gcttcattcg
     1261 cacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1321 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1381 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1441 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     1501 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     1561 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1621 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1681 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1741 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1801 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1861 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1921 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     1981 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2041 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2101 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2161 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2221 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2281 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     2341 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     2401 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
     2461 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     2521 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     2581 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     2641 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     2701 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     2761 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
     2821 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     2881 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     2941 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3001 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3061 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     3121 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3181 agtgtaaagc ctggggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     3241 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3301 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3361 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3421 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3481 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3541 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3601 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3661 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3721 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3781 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3841 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3901 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3961 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4021 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4081 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4141 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4201 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4261 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4321 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4381 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4441 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4501 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4561 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4621 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4681 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4741 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4801 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4861 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4921 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4981 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5041 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5101 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5161 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5221 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     5281 at
//
